Na+ TRANSPORT INHIBITION BY RESPIRATORY SYNCYTIAL VIRUS

呼吸道合胞病毒对 Na 转运的抑制

• 批准号: 6931548
• 负责人: IAN CHRISTOPHER DAVIS
• 金额: $ 5.75万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6931548
• 关键词: Xenopus; gene expression; host organism interaction; ion transport; laboratory mouse; membrane potentials; proteasome; respiratory epithelium; respiratory syncytial virus; sodium ion; tissue /cell culture; ubiquitin; virus infection mechanism; virus protein; voltage /patch clamp

CANDIDATE: I graduated from the University of Bristol in 1992, with Honours in Anatomical Science and Veterinary Science. At UAB I received training in laboratory animal medicine, laboratory animal pathology, and viral pathogenesis in animal models (Ph.D. February 2000; mentor Dr. Patricia Fultz). Subsequently, I elected to join Dr. Matalon's research group, where I might build on my knowledge of viral pathogenesis by cross-mining in electrophysiology, and apply these techniques to study the pathogenesis of human respiratory viral pneumonitides. RESEARCH: Respiratory syncytial virus (RSV) is commonest cause of lower respiratory tract disease in children worldwide. Pathogenesis of RSV-induced bronchiolitis is poorly understood, and effects of RSV infection on ion transport (a seminal function of respiratory epithelial cells) have not been investigated. I hypothesize that RSV infection of respiratory epithelial cells reduces their Na ? transport capacity. Preliminary studies have demonstrated that this hypothesis is correct, both in vitro and in RSV-infeeted BALB/c mice. My aims for years 01- 03 are to: (1) quantify alterations in Na + transport across airway and alveolar epithelia in vivo and ex vivo, after infection of BALB/c mice with RSV; (2) define changes in Na + currents and amiloride-sensitive channel activity after RSV infection of murine epithelial cells in vitro; and (3) correlate alterations in Na + transport induced by RSV in vitro and in vivo with alterations in ENaC expression by marine respiratory epithelia. My plan for years 04-05 is to determine the role of the ubiquitin/proteasome pathway in mediating reduced Na+ transport after RSV infection of respiratory epithelia, and to identify the role of the RSV small hydrophobic (SH) gene product in modulation of Na + transport. I will use a combined electrophysiologic and biochemical approach to investigate effects of RSV on Na + transport at all levels, from the single cell to the whole animal, and to correlate these effects to Na + channel expression and degradation. My project will emphasize cross-training in diverse techniques, including short-circuit measurements across monolayers, radioisotopic ion flux studies, whole cell and single channel patch-clamp, and measurement of alveolar fluid clearance and nasal potential difference in mice. ENVIRONMENT: This SERCA, with Dr. Matalon (lung physiology) as mentor and Dr. Sullender (respiratory syncytial virus) as co-mentor, will provide the training and setting I require for my maturation into an independent scientist focused on comparative pathophysiologic effects of respiratory viruses on normal epithelial cell function.

候选人：我于 1992 年毕业于布里斯托大学，获得解剖学和兽医学荣誉学位。在 UAB，我接受了实验动物医学、实验动物病理学和动物模型病毒发病机制方面的培训（2000 年 2 月获得博士学位；导师 Patricia Fultz 博士）。随后，我选择加入Matalon博士的研究小组，在那里我可以通过电生理学的交叉挖掘来建立我对病毒发病机制的了解，并应用这些技术来研究人类呼吸道病毒性肺炎的发病机制。 研究：呼吸道合胞病毒 (RSV) 是全世界儿童下呼吸道疾病的最常见原因。 RSV 诱发的细支气管炎的发病机制尚不清楚，RSV 感染对离子转运（呼吸道上皮细胞的一种重要功能）的影响尚未得到研究。我假设呼吸道上皮细胞的 RSV 感染会降低其 Na ？运输能力。初步研究表明，无论是在体外还是在感染 RSV 的 BALB/c 小鼠中，这一假设都是正确的。我01年的目标- 03的目的是：（1）在用RSV感染BALB/c小鼠后，体内和离体量化Na + 穿过气道和肺泡上皮细胞转运的变化； (2)定义RSV体外感染小鼠上皮细胞后Na+电流和阿米洛利敏感通道活性的变化； (3) 将体外和体内 RSV 诱导的 Na + 转运的变化与海洋呼吸道上皮细胞 ENaC 表达的变化相关联。我 04-05 年的计划是确定泛素/蛋白酶体途径在 RSV 感染呼吸道上皮后介导 Na+ 转运减少中的作用，并确定 RSV 小疏水 (SH) 基因产物在调节 Na+ 转运中的作用。我将结合使用电生理学和生化学 方法来研究RSV对从单细胞到整个动物的各个水平的Na + 转运的影响，并将这些影响与Na + 通道表达和降解相关联。我的项目将强调多种技术的交叉训练，包括单层短路测量、放射性同位素离子通量研究、全细胞和单通道膜片钳，以及小鼠肺泡液清除率和鼻电位差的测量。 环境：这个 SERCA 以 Matalon 博士（肺生理学）为导师，Sullender 博士（呼吸道合胞病毒）为副导师，将为我提供所需的培训和环境，帮助我成长为一名独立科学家，专注于呼吸道病毒对正常上皮细胞功能的病理生理学影响的比较。